1. Field of the Invention
This invention relates generally to the modulation of neural growth in the central nervous system, and more particularly to methods and associated agents, constructs and compositions for improving CNS neural growth. Specifically, the invention relates to the use of cellular adhesion molecules, and preferably neural cell adhesion molecules such as L1, to foster and improve such neural growth.
2. Description of the Related Art
The ability of neurons to extend neurites is of prime importance in establishing neuronal connections during development. It is also required during regeneration to re-establish connections destroyed as a result of a lesion.
Neurites elongate profusely during development both in the central and peripheral nervous systems of all animal species (Cajal (1928) Degeneration and regeneration in nervous system, Oxford University Press, London). This phenomenon pertains to axons and dendrites. However, in adults, axonal and dendritic regrowth in the central nervous system is increasingly lost with evolutionary progression.
In the peripheral nervous system, after infliction of a lesion, axons of all vertebrate species are able to regrow (Cajal (1928); Martini (1994) J. Neurocytol. 23:1-28). However, in mammals, neurite regrowth following damage is limited to neuritic sprouting. Regrowth of neuronal processes is, however, possible in lower vertebrate species (Stuermer et al. (1992) J. Neurobiol. 23:537-550). In contrast, in the central nervous system, most, if not all neurons of both higher and lower vertebrate adults possess the potential for neurite regrowth (Aguayo (1985) “Axonal regeneration from injured neurons in the adult mammalian central nervous system,” In: Synaptic Plasticity (Cotman, C. W., ed.) New York, The Guilford Press, pp. 457-484.)
Glial cells are the decisive determinants for controlling axon regrowth. Mammalian glial cells are generally permissive for neurite outgrowth in the central nervous system during development (Silver et al. (1982) J. Comp. Neurol. 210:10-29; Miller et al. (1985) Develop. Biol. 111:35-41; Pollerberg et al. (1985) J. Cell. Biol. 101:1921-1929) and in the adult peripheral nervous system (Fawcett et al. (1990) Annu. Rev. Neurosci 13:43-60). Thus, upon infliction of a lesion, glial cells of the adult mammalian peripheral nervous system can revert to some extent to their earlier neurite outgrowth-promoting potential, allowing them to foster regeneration (Kalderon (1988) J. Neurosci Res. 21:501-512; Kliot et al. “Induced regeneration of dorsal root fibres into the adult mammalian spinal cord,” In: Current Issues in Neural Regeneration, New York, pp. 311-328; Carlstedt et al. (1989) Brain Res. Bull. 22:93-102). Glial cells of the central nervous system of some lower vertebrates remain permissive for neurite regrowth in adulthood (Stuermer et al. (1992) J. Neurobiol. 23:537-550). In contrast, glial cells of the central nervous system of adult mammals are not conducive to neurite regrowth following lesions.
Several recognition molecules which act as molecular cues underlying promotion and/or inhibition of neurite growth have been identified (Martini (1996). Among the neurite outgrowth promoting recognition molecules, the neural cell adhesion molecule L1 plays a prominent role in mediating neurite outgrowth (Schachner (1990) Seminars in the Neurosciences 2:497-507). L1-dependent neurite outgrowth is mediated by homophilic interaction. L1 enhances neurite outgrowth on L1 expressing neurites and Schwann cells, and L1 transfected fibroblasts (Bixby et al. (1982) Proc. Nat'l Acad. Sci. U.S.A. 84:2555-2559; Chang et al. (1987) J. Cell. Biol. 104:355-362; Lagenaur et al. (1987) Proc. Natl. Acad. Sci. USA 84:7753-7757; Seilheimer et al. (1988) J. Cell. Biol. 107:341-351; Kadmon et al. (1990a) J. Cell. Biol. 110:193-208; Williams et al. (1992) J. Cell. Biol. 119:883-892). Expression of L1 is enhanced dramatically after cutting or crushing peripheral nerves of adult mice (Nieke et al. (1985) Differentiation 30:141-151; Martini et al. (1994a) Glia 10:70-74). Within two days L1 accumulates at sites of contact between neurons and Schwann cells being concentrated mainly at the cell surface of Schwann cells but not neurons (Martini et al. (1994a)). Furthermore, the homophilic binding ability of L1 is enhanced by molecular association with the neural cell adhesion molecule N-CAM, allowing binding to occur through homophilic assistance (Kadmon et al. (1990a); Kadmon et al. (1990b) J. Cell Biol. 110:209-218 and 110:193-208; Horstkorte et al. (1993) J. Cell. Biol. 121:1409-1421). Besides its neurite outgrowth promoting properties, L1 also participates in cell adhesion (Rathjen et al. (1984) EMBO J. 3:1-10; Kadmon et al. (1990b) J. Cell. Biol. 110:209-218; Appel et al. (1993) J. Neurosci., 13:4764-4775), granule cell migration (Lindner et al. (1983) Nature 305:427-430) and myelination of axons (Wood et al. (1990) J. Neurosci 10:3635-3645).
L1 consists of six immunoglobulin-like domains and five fibronectin type III homologous repeats. L1 acts as a signal transducer, with the recognition process being a first step in a complex series of events leading to changes in steady state levels of intracellular messengers. The latter include inositol phosphates, Ca2+, pH and cyclic nucleotides (Schuch et al. (1990) Neuron 3:13-20; von Bohlen und Hallbach et al. (1992) Eur. J. Neurosci. 4:896-909; Doherty et al. (1992) Curr. Opin. Neurobiol. 2:595-601) as well as changes in the activities of protein kinases such as protein kinase C and pp60c-src (Schuch et al. (1990) Neuron 3:13-20; Atashi et al. (1992) Neuron 8:831-842). L1 is also associated with a casein type II kinase and another unidentified kinase which phosphorylates L1 (Sadoul et al. (1989) J. Neurochem 328:251-254). L1-mediated neurite outgrowth is sensitive to the blockage of L type Ca2+ channels and to pertussis toxin. These findings indicate the importance of both Ca2+ and G proteins in L1-mediated neurite outgrowth (Williams et al. (1992) J. Cell. Biol. 119:883-892). L1 is also present on proliferating, immature astrocytes in culture and neurite outgrowth is promoted on these cells far better than on differentiated, L1 immunonegative astrocytes (Saad et al. (1991) J. Cell. Biol. 115:473-484). In vivo, however, astrocytes have been found to express L1 at any of the developmental stages examined from embryonic day 13 until adulthood (Bartsch et al. (1989) J. Comp. Neurol 284:451-462; and unpublished data).
Given the capability of L1 to promote neurite outgrowth, it is pertinent to investigate whether astrocytic expression of L1 and other members of the immunoglobulin superfamily to which L1 belongs, may overcome potentially inhibitory molecular cues reported to be present on glial cells and myelin in the adult central nervous system (Schachner et al., Perspectives in Developm. Neurobiol. in Press; Schwab et al. (1993) Ann. Rev. Neurosci. 16:565-595). This is of particular relevance to the development of effective strategies for the treatment of debilitation caused by the malformation of or injury to neural tissues of the CNS, and it is toward such objectives that the present invention is directed.